A multiplexed ARMS-PCR approach for the detection of common MECP2 mutations

Genet Test Mol Biomarkers. 2009 Feb;13(1):19-22. doi: 10.1089/gtmb.2008.0051.

Abstract

Rett syndrome (RTT), an X-linked dominant neurodevelopmental disorder, is caused mainly by de novo mutations in the methyl-CpG-binding protein 2 gene (MECP2). Although more than 200 different MECP2 mutations have been identified throughout the gene, 7 of those (p.R133C, p.T158M, p.R168X, p.R255X, p.R270X, p.R294X, and p.R306C) account for up to two-thirds of pathogenic mutations in RTT patients. A rapid and efficient screening strategy for these mutations can be used as a preliminary step for genetic diagnosis of RTT. The current protocols used for this purpose are of high cost and require special equipment. We have designed a simpler multiplex amplification refractory mutation system (ARMS)-PCR strategy that allows identification of these common MECP2 mutant alleles in four PCR reactions. The assay was tested in 14 RTT patients who were previously genotyped using PCR-restriction fragment length polymorphism and DNA sequencing. A complete concordance was observed between the results of the two methods. The multiplex ARMS-PCR does not require any special equipment, and it provides rapid, reproducible, and cost-effective detection of common MECP2 mutations. The assay can be carried out efficiently in a standard molecular genetics laboratory and suitable as a preliminary screen for all patients with RTT diagnosis.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • Base Sequence
  • DNA Mutational Analysis / methods*
  • DNA Primers / genetics
  • Female
  • Humans
  • Male
  • Methyl-CpG-Binding Protein 2 / genetics*
  • Point Mutation
  • Polymerase Chain Reaction / methods*
  • Polymorphism, Restriction Fragment Length
  • Rett Syndrome / diagnosis
  • Rett Syndrome / genetics*

Substances

  • DNA Primers
  • MECP2 protein, human
  • Methyl-CpG-Binding Protein 2